The published data also support the hypothesis that increased VLA

The published data also support the hypothesis that increased VLA-4 will allow for improved in vivo function and improved ability to accumulate within the granuloma. One could propose therefore that the level of nitric oxide within the lesional site can dramatically impact the local protective and immunopathological response by reducing accumulation of specific subsets of activated effector cells and by altering the potency of the lymphocytes with regard to accumulation within the lesion and cytokine production. By demonstrating the differential impact of nitric oxide on distinct check details functional subsets of cells, we have identified a mechanism whereby

protection and pathology in mycobacterial disease are modulated by nitric oxide. The development of inflammation during mycobacterial infection is an important component of the disease process and is actively modulated by CD4+ T cells. Herein, we demonstrate that within the pool of effector T cells, there is an activated T-cell subset that is more selleck inhibitor susceptible to the regulatory factors active within the granuloma. Defining the relative protective and pathological role of this activated T-cell subset (CD4+T-bet+CD69loVLA4hi) will allow for improved vaccination and immunotherapeutic intervention. All mice were bred

at the Trudeau Institute and were treated according to National Institutes of Health and Trudeau Institute Animal Care and Use Committee guidelines. All animal protocols used herein were approved by the Trudeau Institute Animal Care and Use Committee. C57BL/6 and B6.129P2-nos2tm1Lau (nos2−/−) (originally purchased from JAX Mice, Maine) OSBPL9 were used in these experiments. Mice were infected with M. avium 25291 (ATTC) at a dose of 1 × 106 cfu by lateral tail vein injection. The level of bacteria in specific organs was determined by homogenizing the organs and plating on agar and counting colonies [48, 49]. Some infected WT mice were treated with aminoguanidine hemisulfate (Sigma-Aldrich) at 2.5 g/100 mL in the drinking water for defined periods of time; control mice received water without drug.

Liver sections were placed in 10% neutral-buffered formalin, blocked in paraffin, processed for light microscopy, and stained with hematoxylin and eosin to provide cell structure. For immunofluorescence staining, liver sections were harvested into cold HBSS and 3–4 mm sections cut with a scalpel. Sections were placed in 4% low-melt agarose (Lonza Seaplaque Agarose, Fisher Scientific) in HBSS. The solidified gel containing sections of liver was then sectioned using a vibrating microtome cooled to 4°C (Leica VT1000) and 200-micron sections were collected into 12-well plates containing HBSS, FcBlock, 5% normal mouse serum. Sections were stained with fluorescently labeled antibodies, anti-CD4 PE (RM4–5), anti-CD8 PE (clone 53–6.

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